항체발현 파지기법을 이용한 B형 간염바이 러스 표면항원 preS1과 preS2에 대한 인간 항체의 생산

다국어 초록 (Multilingual Abstract)

Human monoclonal antibodies have considerable potential in the prophylaxis and treatment of viral disease. By cloning human ig gene segments from the B cells of volunteer into pComb3 phagemid vector, antibody library was created of filamentous phage particle displaying Fab fragments on their surface after rescuing with M13K07 helper phage. The size of library was 7×10^9 pfu. Phage antibodies(phabs) were panned against biotinylated preS1 using streptavidine coated Dynabead. The soluble Fab antibodies were prepared from phagemid colonies and assayed directly for the ability to bind preS1 by ELISA. And then 3DW and 8GW specific to preS1 which have both heavy and light chain to form Fab fragment, were selected. The soluble Fab antibody from 3DW was expressed highly at the concentration of 0.1 - 10. mM of IPTG, and 5 hours postinduction The soluble antibodies from 3DW and 8GW showed their relative affinities of 2×10^6 M^-1, and 5×10^7 M^-1, respectively, and the specificities to preS1 on ELISA. The relative affinities of soluble Fab antibodies from G23W and B66W were 3×10^6 M^-1와 5×10^6 M^-1, respectively and theses antibodies were specific to preS2 and its related protein.
Our results suggest that antibody phage display library is very useful method to generate the human monoclonal antibody and that the human Fab monoclonal antibodies specific to preS1 and preS2 selected in this study open the way to treat hepatitis B as a component of passive immunotherapeutics.

목차 (Table of Contents)

• Abstract
• 서론
• 재료 및 방법
• 결과
• 고찰
• 인용문헌